Scientists use gene editing to eliminate viruses in live pigs

First Pigs without Active Porcine Endogenous Retrovirus (PERV) Produced Using CRISPR-Cas9 Genome Editing Technology – a Landmark Advance for Xenotransplantation

Scientists have edited the pig genome to deactivate a family of retroviruses. The results hold important implications for transplant medicine in humans. The shortage of human organs and tissues for transplantation represents one of the most significant unmet medical needs.

One promising prospect is to use animal organs in humans, with pig organs being particularly compatible for such transplantation. The pig genome, however, includes porcine endogenous retroviruses (PERVs), which can be passed on to other cells when cultured together.

Gene editing techniques could prove useful for removing virus genes from the pig genome, paving the way for pig-to-human transplants, yet efforts have so far only been successfully demonstrated in cell lines, not live animals. Here, George Church, Dong Niu and colleagues demonstrate the feat in live animals. eGenesis, a biotechnology company co-founded by George church, focused on transforming xenotransplantation into a lifesaving medical procedure, announced the publication of the study in the journal Science.

eGenesis scientists and their collaborators demonstrated the inactivation of PERV to prevent cross-species viral transmission and a breakthrough in producing the first PERV-free pigs, an important milestone for xenotransplantation.

The team first confirmed that PERVs in pig cells can be transmitted to human cells when cultured together. Exposing human cells infected with PERV to uninfected humans cells also resulted in transmission, highlighting the need to deactivate PERVs in pigs if transplantation is to one day, occur.

Next, the researchers mapped and characterized the PERVs present in the genome of pig fibroblast cells, identifying 25 in total. They used the gene editing tool CRISPR to deactivate all 25 genomic sites. Despite the presence of highly modified cells in the population, none of the cloned cells could be grown with greater than 90% PERV editing efficiency. By adding a concoction of additional factors related to DNA repair, however, the team was able to grow viable cells with 100% of PERVs deactivated.

When they implanted the embryos into sows, they found that the resulting piglets exhibited no signs of PERVs, with some piglets surviving up to four months after birth.

“This is the first publication to report on PERV-free pig production,” said Luhan Yang, Ph.D., co-founder and chief scientific officer at eGenesis. “We generated a protocol to enable multiplex genome editing, eradicated all PERV activity using CRISPR technology in cloneable primary porcine fibroblasts and successfully produced PERV-free piglets. This research represents an important advance in addressing safety concerns about crossspecies viral transmission. Our team will further engineer the PERV-free pig strain to deliver safe and effective xenotransplantation.”

Co-founder, Author and Editor, biotechin.asia, Biotech Media Pte. Ltd.
Laxmi graduated from University of Mumbai, India (Bachelors in Biotechnology and Biochemistry and Masters in Biochemistry) in 2007 and received her Ph.D in Virology from Nanyang Technological University, Singapore. She worked on the process of assembly of respiratory syncytial virus in macrophages and epithelial cells and has several papers to her credit.